This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are proposing studies to determine the resolution limits for fluorescence diffuse optical tomography (F-DOT) using over-sampled measurements. It will exploit the combined expertise of two NCRR National Resources: data acquisition using the recently established molecular imaging workbench at the Duke Center for In Vivo Microscopy (CIVM) and image reconstruction using the parallel computing facilities at the Pittsburgh Supercomputing Center (PSC). Specifically, we will be conducting the following studies: (i) Development of open-structured software package for F-DOT simulations and image reconstruction;(ii) Study of resolution limit in phantoms and animals without prior information;and (iii) Study the improvement of resolution limit with simultaneously acquired prior information from micro-CT.